Molecular diagnosis of genetic defects and diseases requires techniques that are capable of detecting minute quantities of DNA and RNA in a sample, or techniques that are extremely sensitive to detect mutations in DNA. For example, techniques such as southern blot, polymerase chain reaction, reverse transcriptase-polymerase chain reaction and ligase chain reaction have been extensively used to detect microbial and viral pathogens, such as HIV, and to diagnosis cancers and genetic diseases, such as cystic fibrosis and muscular dystrophy. Specifically, techniques such as polymerase chain reaction, or PCR, amplify small quantities of the target nucleotide sequence to obtain detectable quantities.
However, techniques such as PCR typically require additional separation procedures prior to detection of the target sequence to achieve adequate sensitivity. Moreover, PCR has a high rate of sample-to-sample contamination which decreases the accuracy of the procedure.
The development of simple, fast and reliable amplification-based assays to detect target nucleic acids will greatly aid molecular diagnosis.